Subunit nanovaccine elicited T cell functional activation controls Trypanosoma cruzi mediated maternal and placental tissue damage and improves pregnancy outcomes in mice.

This study investigated a candidate vaccine effect against maternal Trypanosoma cruzi (Tc) infection and improved pregnancy outcomes. For this, TcG2 and TcG4 were cloned in a nanoplasmid optimized for delivery, antigen expression, and regulatory compliance (nano2/4 vaccine). Female C57BL/6 mice were immunized with nano2/4, infected (Tc SylvioX10), and mated 7-days post-infection to enable fetal development during the maternal acute parasitemia phase. Females were euthanized at E12-E17 (gestation) days. Splenic and placental T-cell responses were monitored by flow cytometry. Maternal and placental/fetal tissues were examined for parasites by qPCR and inflammatory infiltrate by histology. Controls included age/immunization-matched non-pregnant females. Nano2/4 exhibited no toxicity and elicited protective IgG2a/IgG1 response in mice. Nano2/4 signaled a splenic expansion of functionally active CD4+ effector/effector memory (Tem) and central memory (Tcm) cells in pregnant mice. Upon challenge infection, nano2/4 increased the splenic CD4+ and CD8+T cells in all mice and increased the proliferation of CD4+Tem, CD4+Tcm, and CD8+Tcm subsets producing IFNγ and cytolytic molecules (PRF1, GZB) in pregnant mice. A balanced serum cytokines/chemokines response and placental immune characteristics indicated that pregnancy prevented the overwhelming damaging immune response in mice. Importantly, pregnancy itself resulted in a significant reduction of parasites in maternal and fetal tissues. Nano2/4 was effective in arresting the Tc-induced tissue inflammatory infiltrate, necrosis, and fibrosis in maternal and placental tissues and improving maternal fertility, placental efficiency, and fetal survival. In conclusion, we show that maternal nano2/4 vaccination is beneficial in controlling the adverse effects of Tc infection on maternal health, fetal survival, and pregnancy outcomes.

Acute muscle mass loss was alleviated with HMGB1 neutralizing antibody treatment in severe burned rats.

Burn injury is associated with muscle wasting, though the involved signaling mechanisms are not well understood. In this study, we aimed to examine the role of high mobility group box 1 (HMGB1) in signaling hyper-inflammation and consequent skeletal muscle impairment after burn. Sprague Dawley rats were randomly assigned into three groups: (1) sham burn, (2) burn, (3) burn/treatment. Animals in group 2 and group 3 received scald burn on 30% of total body surface area (TBSA) and immediately treated with chicken IgY and anti-HMGB1 antibody, respectively. Muscle tissues and other samples were collected at 3-days after burn. Body mass and wet/dry weights of the hind limb muscles (total and individually) were substantially decreased in burn rats. Acute burn provoked the mitochondrial stress and cell death and enhanced the protein ubiquitination and LC3A/B levels that are involved in protein degradation in muscle tissues. Further, an increase in muscle inflammatory infiltrate associated with increased differentiation, maturation and proinflammatory activation of bone marrow myeloid cells and αß CD4+ T and γδ T lymphocytes was noted in in circulation and spleen of burn rats. Treatment with one dose of HMGB1 neutralizing antibody reduced the burn wound size and preserved the wet/dry weights of the hind limb muscles associated with a control in the markers of cell death and autophagy pathways in burn rats. Further, anti-HMGB1 antibody inhibited the myeloid and T cells inflammatory activation and subsequent dysregulated inflammatory infiltrate in the muscle tissues of burn rats. We conclude that neutralization of HMGB1-dependent proteolytic and inflammatory responses has potential beneficial effects in preventing the muscle loss after severe burn injury.

Effects of Acute and Chronic Trypanosoma cruzi Infection on Pregnancy Outcomes in Mice: Parasite Transmission, Mortality, Delayed Growth, and Organ Damage in Pups.

Chagas disease is caused by Trypanosoma cruzi. This study aimed to determine the effects of T. cruzi infection on fertility rate and health of the newborn pups in pregnant mice. Female mice were challenged with T. cruzi and mated at 21 days (acute parasitemic phase) or 90 days (chronic parasite persistence phase) after infection. Pups were examined for growth up to 20 days after birth; and parasite burden in brain, heart, skeletal muscle, and intestine was measured by real-time quantitative PCR. The inflammatory infiltrate, necrosis, and fibrosis in pups' heart and brain tissues were evaluated by histology. T. cruzi infection in dams delayed the onset of pregnancy, decreased the fertility rate, and led to vertical transmission of parasite to the pups. Furthermore, infected dams delivered pups that exhibited decreased survival rate, decreased birth weight, and decreased growth rate. Significantly increased inflammation, necrosis, and fibrosis of cardiac and brain tissues were noted in pups born to infected dams. Initial challenge with higher parasite dose had more detrimental effects on fertility rate and pups' health in both acutely and chronically infected dams. In conclusion, mice offer a promising model to evaluate the efficacy of new vaccines and therapeutic drugs in controlling the acute and chronic maternal T. cruzi infection and congenital transmission to newborns, and in improving the fertility rate and pups' health outcomes.

Animal Models of Trypanosoma cruzi Congenital Transmission.

Chagas disease, initiated by the etiological agent Trypanosoma cruzi, is an endemic infection in the American continent. Although vectorial transmission of T. cruzi is recognized as the main mode of infection, other routes such as congenital and blood transfusion are also documented as important methods of transmission. T. cruzi maternal-fetal transmission has been recorded in humans and examined by some investigators in naturally and experimentally infected mammals. Dogs are recognized as the major reservoir host in maintaining the domestic transmission of T. cruzi; however, the importance of congenital transmission in preserving the infection cycle in dogs has not been studied in detail. In this article, we reviewed the current knowledge of congenital transmission of T. cruzi in humans and compared the placental architecture of humans and different animals with particular attention to rodents, dogs, and non-human primates that have been used as experimental models of T. cruzi infection, congenital transmission, and Chagas disease pathogenesis. The placentas of humans and animals have some similar and dissimilar characteristics that should inform the study design and interpretation of results when evaluating the efficacy of new anti-parasite drugs and therapies against congenital infection.

Oxidative stress implications for therapeutic vaccine development against Chagas disease.

INTRODUCTION: Pathogenesis of Chagas disease (CD) caused by the protozoan parasite Trypanosoma cruzi (T. cruzi) involves chronic oxidative and inflammatory stress. In this review, we discuss the research efforts in therapeutic vaccine development to date and the potential challenges imposed by oxidative stress in achieving an efficient therapeutic vaccine against CD. AREAS COVERED: This review covers the immune and nonimmune mechanisms of reactive oxygen species production and immune response patterns during T. cruzi infection in CD. A discussion on immunotherapy development efforts, the efficacy of antigen-based immune therapies against T. cruzi, and the role of antioxidants as adjuvants is discussed to provide promising insights to developing future treatment strategies against CD. EXPERT OPINION: Administration of therapeutic vaccines can be a good option to confront persistent parasitemia in CD by achieving a rapid, short-lived stimulation of type 1 cell-mediated immunity. At the same time, adjunct therapies could play a critical role in the preservation of mitochondrial metabolism and cardiac muscle contractility in CD. We propose combined therapy with antigen-based vaccine and small molecules to control the pathological oxidative insult would be effective in the conservation of cardiac structure and function in CD.

Autophagy regulator Atg9 is degraded by the proteasome.

Autophagy is a highly conserved biological process essential to protein, cellular and organismal homeostasis. As autophagy plays a critical role in cellular responses to various external and internal stimuli, it is important to understand the mechanism underlying autophagy regulation. Here, we monitor the stability of 17 key autophagy factors in the yeast S. cerevisiae and show that Atg9 and Atg14 are degraded under normal growth conditions. Whereas Atg14 is regulated by both the proteasome and autophagy, Atg9 turnover is normally mediated by the proteasome but impeded upon starvation or rapamycin treatment. Interestingly, distinct segments of Atg9 confer instability, suggesting that multiple pathways are involved in Atg9 degradation. Our results provide the foundation to further elucidate the physiological significance of Atg9 turnover and also the interplay between two major proteolytic systems (i.e., autophagy and the proteasome).

Epidemiology and pathogenesis of maternal-fetal transmission of Trypanosoma cruzi and a case for vaccine development against congenital Chagas disease.

Trypanos o ma cruzi (T. cruzi or Tc) is the causative agent of Chagas disease (CD). It is common for patients to suffer from non-specific symptoms or be clinically asymptomatic with acute and chronic conditions acquired through various routes of transmission. The expecting women and their fetuses are vulnerable to congenital transmission of Tc. Pregnant women face formidable health challenges because the frontline antiparasitic drugs, benznidazole and nifurtimox, are contraindicated during pregnancy. However, it is worthwhile to highlight that newborns can be cured if they are diagnosed and given treatment in a timely manner. In this review, we discuss the pathogenesis of maternal-fetal transmission of Tc and provide a justification for the investment in the development of vaccines against congenital CD.

Immunity and vaccine development efforts against Trypanosoma cruzi.

Trypanosoma cruzi (T. cruzi) is the causative agent for Chagas disease (CD). There is a critical lack of methods for prevention of infection or treatment of acute infection and chronic disease. Studies in experimental models have suggested that the protective immunity against T. cruzi infection requires the elicitation of Th1 cytokines, lytic antibodies and the concerted activities of macrophages, T helper cells, and cytotoxic T lymphocytes (CTLs). In this review, we summarize the research efforts in vaccine development to date and the challenges faced in achieving an efficient prophylactic or therapeutic vaccine against human CD.

[Stunting, overweight and obesity during the nutrition transition in schoolchildren of Arandas, Jalisco, Mexico]. / Desmedro, sobrepeso y obesidad durante la transición nutricia en escolares de Arandas, Jalisco, México.

BACKGROUND: Nutrition transition provokes changes in the nutritional status of individuals subjected to the interaction of various environmental factors; therefore, the aim was to demonstrate that nutrition transition is associated with socioeconomic changes, eating habits and physical activity, potentially involved in stunting, overweight and obesity of schoolchildren. METHODS: Case-control study. 102 participants from the Instituto Alteño para el Desarrollo de Jalisco (cases) and 194 from the elementary school system (controls), aged 5 to 12 years, were included. Dependent variables were these indexes weight/age (Z), height/age (Z), BMI (Z). Independent variables were the socioeconomic and demographic characteristics. Student's t test, chi square, odds ratio (OR) and 95% confidence intervals (95% CI) were estimated. RESULTS: Family income was low (p = 0.031) and unstable job was higher in cases: OR = 4.1, 95% CI = 2.8-6.0. The frequency of stunting was higher in cases (9.9% vs. 5.9%). The combination of overweight/obesity was higher in controls (27.3% vs. 16.8%), OR = 1.85, 95% CI = 1.0-3.4. CONCLUSIONS: The nutritional status of children of Arandas, Jalisco, has been modified by an accelerate nutrition transition, provoked by socioeconomic, educational and demographic factors that might have influence on the persistence of stunting and an increasing prevalence of overweight/obesity.

INTRODUCCIÓN: la transición nutricia (TN) propicia cambios en el estado de nutrición de los individuos sometidos a la interacción de diversos factores ambientales; por tanto, el objetivo fue demostrar que la TN se asocia a cambios socioeconómicos, en los hábitos de alimentación y en la actividad física, potencialmente implicados en el desmedro, el sobrepeso y la obesidad en escolares. MÉTODOS: estudio de casos (n = 102) del Instituto Alteño para el Desarrollo de Jalisco y controles (n = 194) de escuelas públicas del municipio, de cinco a doce años de edad. Las variables dependientes fueron los índices peso/edad (Z), talla/edad (Z), IMC/E (Z). Las variables independientes fueron las características socioeconómicas y demográficas. Se utilizaron las pruebas t de Student, chi cuadrada y razón de momios (RM) con intervalos de confianza al 95% (IC 95%). RESULTADOS: el ingreso familiar mensual fue menor (p = 0.031) y el trabajo eventual fue más común en los casos, RM = 4.1, IC 95% = 2.8-6.0. La frecuencia de desmedro fue mayor en los casos (9.9% frente a 5.9%). La combinación de sobrepeso/obesidad fue mayor en los controles (27.3% frente a 16.8%), RM = 1.85, IC 95% = 1.0-3.4. CONCLUSIÓN: el estado nutricio de los niños de Arandas parece estar modificándose por una transición nutricia acelerada por cambios socioeconómicos, demográficos y educacionales que influyen en la persistencia de desmedro y en una creciente prevalencia de sobrepeso y obesidad.

[Factors associated with physical activity and body mass index among schoolchildren from Arandas, Jalisco, Mexico]. / Factores asociados a la actividad física y el índice de masa corporal en escolares de Arandas, Jalisco, México.

BACKGROUND: In 2015, Mexico was the top country for childhood obesity. The objective was to identify the association between physical activity and sedentary lifestyle with the social and demographic characteristics of families of schoolchildren in Arandas, Jalisco, Mexico. METHODS: In a cross-sectional study, 192 schoolchildren were randomly selected. Body mass index (BMI), active and sedentary behaviors, and socio-demographic characteristics of families were obtained. Logistic regression models with the included variables were constructed. RESULTS: Males living in not-crowding houses [OR 6.12 (2.17-17.25), p = 0.001], whose mothers were housewives [OR 2.44 (1.00, 5.94), p = 0.05], practiced more physical activity. Active transport to school was more common in schoolchildren whose fathers had lower income [OR 3.13 (1.27, 7.7), p = 0.013] and employment as peasant or mason [OR 5.12 (1.13, 23.3), p = 0.034]. Schoolchildren of nuclear families spent more hours watching television [OR 2.69 (1.10, 6.58), p = 0.03]. The frequency of outdoor playing was higher in males whose fathers had unstable employment [OR 2.93 (1.06, 8.1), p = 0.038] and low education [OR 2.94 (0.96, 8.98), p = 0.059]. CONCLUSION: Families with lower socioeconomic strata (less educated parents, unstable employment and family overcrowding) are more associated with active activities that do not require active economic spending.

Introducción: en 2015, México ocupaba el primer lugar mundial en obesidad infantil. El objetivo fue identificar la asociación entre conductas activas y sedentarias con características sociodemográficas de la familia en escolares de Arandas, Jalisco, México. Métodos: en estudio transversal se seleccionaron aleatoriamente 192 escolares. Se obtuvieron el índice de masa corporal (IMC), las conductas activas y sedentarias, y las características sociodemográficas de las familias. Se construyeron modelos de regresión logística con las variables incluidas. Resultados: los varones en condiciones de no hacinamiento (razón de momios [RM] 6.12, intervalo de confianza al 95% [IC 95%] 2.17-17.25, p = 0.001) cuyas madres se dedicaban al hogar (RM 2.44, IC 95% 1.00-5.94, p = 0.05) refirieron que practicaban actividad física con más frecuencia. El transporte activo hacia la escuela fue más común en hijos de padres con menor ingreso económico (RM 3.13, IC 95% 1.27-7.7, p = 0.013) y con empleo de albañil o campesino (RM 5.12, IC 95% 1.13-23.3, p = 0.034). Los escolares provenientes de familias nucleares pasaron más horas frente al televisor (RM 2.69, IC 95% 1.10-6.58, p = 0.03). La frecuencia de juegos en la calle fue significativamente mayor en varones cuyos papás tuvieron un empleo inestable (RM 2.93, IC 95% 1.06-8.1, p = 0.038) y baja escolaridad (RM 2.94, IC 95 % 0.96-8.98, p = 0.059). Conclusión: las familias con estrato socioeconómico más bajo se asociaron con mayor frecuencia a actividades activas que no requieren un gasto económico.

Risk factors associated with iron depletion and parasites in preschool and school children of Arandas, Jalisco, México / Factores de riesgo asociados con la depleción de hierro y parasitosis en niños preescolares y escolares de Arandas, Jalisco, México

Objective: The objective of this study is to explore factors associated with iron depletion and parasites in preschool and school children attending the Instituto Alteño para el Desarrollo de Jalisco (INADEJ) and children who attend preschools and schools in Arandas, Jalisco. Methods: This cross sectional study assessed two groups of children. In the INADEJ group, 102 children aged 60 to 144 months old were included, and in the School group of Arandas, Jalisco, 206 children were randomly selected from schools and preschools in the same municipality. Variables: Ferritin concentration (μg/ml), parasites, demographic, socioeconomic, educational and dietary data were collected. The Student’s t test, U Mann Whitney, chi square, odds ratio and logistic regressions were estimated. Results: Family income in the Schools group was $5,707 Mexican pesos (MP), which was higher than the family income in the INADEJ group ($4,311 MP), p = 0.031. The proportion of parents with incomplete primary education was higher in the INADEJ group (41.3%) than in the Schools group (35.4%) [OR = 1.88 (1.0-3.55)]. Unstable work was more frequent in the INADEJ group than in the Schools group [OR = 5.6 (3.26-9.62)]. The ferritin concentration was lower in the INADEJ group than in the Schools group (25 μg/L vs. 60 μg/mL, respectively), p <0.001. Having giardiasis and two or more parasites was more common in the INADEJ group compared with the Schools group [OR = 7.2 (2.25-23.0)] and [RM 3.86 (1.35- 10.98)]. Heme iron intake was lower in the INADEJ group. Conclusion: Low family income, father’s job instability, lower levels of education, overcrowding, living with animals and lower consumption of heme iron were associated with parasites and iron depletion (AU)

Objetivo: Explorar factores asociados con depleción de hierro y parasitosis en preescolares y escolares que asisten al Instituto Alteño para el Desarrollo de Jalisco (INADEJ) y los niños que asisten a escuelas en Arandas, Jalisco. Métodos: Este estudio transversal evaluó dos grupos de niños. En el grupo INADEJ, fueron incluidos 102 niños de 60 a 144 meses de edad; en el grupo Escuelas fueron seleccionados 206 niños al azar de las escuelas del mismo municipio. Se obtuvieron las variables: ferritina (g/mL), parasitosis, variables demográficas, socioeconómicas, educativas y datos dietéticos. Se realizaron pruebas t de Student, U de Mann-Whitney, ji cuadrada; odds ratio y regresiones logísticas. Resultados: El ingreso familiar en el grupo escuelas era $ 5,707 pesos mexicanos (pm), más alto que el grupo INADEJ ($ 4,311 pm), p=0,031. La proporción de padres con educación primaria incompleta fue mayor en el grupo INADEJ (41,3%) que en el grupo escuelas (35,4%) [OR = 1,88 (1,0-3,55)]. Trabajo inestable del padre fue más frecuente en el grupo INADEJ [OR = 5,6 (3,26-9,62)]. La concentración de ferritina fue menor en el grupo INADEJ que en el grupo Escuelas (25 μg/L vs. 60 μg/ml, respectivamente), p <0,001. Tener giardiasis y dos o más parásitos fue más frecuente en el grupo INADEJ [OR = 7,2 (2,25 a 23,0)] y [RM 3,86 (1,35-10,98)]. La ingestión de hierro heme fue menor en el grupo INADEJ. Conclusion: Los bajos ingresos familiares, inestabilidad laboral de su padre, pobre educación, hacinamiento, convivencia con animales y menor consumo de hierro hemo se asociaron a parasitosis y depleción de hierro de hierro (AU)

Risk factors associated with iron depletion and parasites in preschool and school children of Arandas, Jalisco, México.

OBJECTIVE: The objective of this study is to explore factors associated with iron depletion and parasites in preschool and school children attending the Instituto Alteño para el Desarrollo de Jalisco (INADEJ) and children who attend preschools and schools in Arandas, Jalisco. METHODS: This cross sectional study assessed two groups of children. In the INADEJ group, 102 children aged 60 to 144 months old were included, and in the School group of Arandas, Jalisco, 206 children were randomly selected from schools and preschools in the same municipality. VARIABLES: Ferritin concentration (µg/ml), parasites, demographic, socioeconomic, educational and dietary data were collected. The Student's t test, U Mann Whitney, chi square, odds ratio and logistic regressions were estimated. RESULTS: Family income in the Schools group was $5,707 Mexican pesos (MP), which was higher than the family income in the INADEJ group ($4,311 MP), p = 0.031. The proportion of parents with incomplete primary education was higher in the INADEJ group (41.3%) than in the Schools group (35.4%) [OR = 1.88 (1.0-3.55)]. Unstable work was more frequent in the INADEJ group than in the Schools group [OR = 5.6 (3.26-9.62)]. The ferritin concentration was lower in the INADEJ group than in the Schools group (25 µg/L vs. 60 µg/mL, respectively), p <0.001. Having giardiasis and two or more parasites was more common in the INADEJ group compared with the Schools group [OR = 7.2 (2.25-23.0)] and [RM 3.86 (1.35-10.98)]. Heme iron intake was lower in the INADEJ group. CONCLUSION: Low family income, father's job instability, lower levels of education, overcrowding, living with animals and lower consumption of heme iron were associated with parasites and iron depletion.

Objetivo: Explorar factores asociados con depleción de hierro y parasitosis en preescolares y escolares que asisten al Instituto Alteño para el Desarrollo de Jalisco (INADEJ) y los niños que asisten a escuelas en Arandas, Jalisco. Métodos: Este estudio transversal evaluó dos grupos de niños. En el grupo INADEJ, fueron incluidos 102 niños de 60 a 144 meses de edad; en el grupo Escuelas fueron seleccionados 206 niños al azar de las escuelas del mismo municipio. Se obtuvieron las variables: ferritina (g/mL), parasitosis, variables demográficas, socioeconómicas, educativas y datos dietéticos. Se realizaron pruebas t de Student, U de Mann-Whitney, ji cuadrada; odds ratio y regresiones logísticas. Resultados: El ingreso familiar en el grupo escuelas era $ 5,707 pesos mexicanos (pm), más alto que el grupo INADEJ ($ 4,311 pm), p=0,031. La proporción de padres con educación primaria incompleta fue mayor en el grupo INADEJ (41,3%) que en el grupo escuelas (35,4%) [OR = 1,88 (1,0-3,55)]. Trabajo inestable del padre fue más frecuente en el grupo INADEJ [OR = 5,6 (3,26-9,62)]. La concentración de ferritina fue menor en el grupo INADEJ que en el grupo Escuelas (25 µg/L vs. 60 µg/ml, respectivamente), p.